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T3SSs are widely distributed virulence determinants of Gram-negative bacteria. They translocate bacterial proteins into host cells to manipulate them during infection. Only upon direct physical contact with host cells are T3SSs activated to secrete their effectors of virulence. T3SSs are composed of a cytoplasmic secretion machinery and a region spanning both bacterial membranes into which a hollow needle is embedded, protruding from the bacterial surface. The needle is the secretion channel and also directly involved in regulating secretion, probably by transmitting the host cell-detection signal. The needle distal tip seems to be the host cell sensor and then forms the translocation pore within the host membrane. We here aim to understand the detailed structure of the needle tip before and during host cell sensing and translocon insertion so as to be able to deduce how it structurally senses, transmits and responds to the host-cell contact signal. This can be broken down into: Vis ualisation of the Shigella tip complex (TC) in different functional states; Determination of the precise arrangement of components in the resting TC; Development of a highly detailed view of how the TC interacts with the needle.
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